Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degradation in Bacillus subtilis.

Detalhes bibliográficos
Autor(a) principal: Sá-Nogueira, Isabel de
Data de Publicação: 2008
Outros Autores: Inácio, José Manuel, Correia, Isabel Lopes
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/4200
Resumo: Microbiology 154 (2008) 2719-2729
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spelling Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degradation in Bacillus subtilis.Microbiology 154 (2008) 2719-2729Bacillus subtilis produces alpha-l-arabinofuranosidases (EC 3.2.1.55; AFs) capable of releasing arabinosyl oligomers and l-arabinose from plant cell walls. Here, we show by insertion-deletion mutational analysis that genes abfA and xsa(asd), herein renamed abf2, encode AFs responsible for the majority of the intracellular AF activity in B. subtilis. Both enzyme activities were shown to be cytosolic and functional studies indicated that arabino-oligomers are natural substrates for the AFs. The products of the two genes were overproduced in Escherichia coli, purified and characterized. The molecular mass of the purified AbfA and Abf2 was about 58 kDa and 57 kDa, respectively. However, native PAGE gradient gel analysis and cross-linking assays detected higher-order structures (>250 kDa), suggesting a multimeric organization of both enzymes. Kinetic experiments at 37 degrees C, with p-nitrophenyl-alpha-l-arabinofuranoside as substrate, gave an apparent K(m) of 0.498 mM and 0.421 mM, and V(max) of 317 U mg(-1) and 311 U mg(-1) for AbfA and Abf2, respectively. The two enzymes displayed maximum activity at 50 degrees C and 60 degrees C, respectively, and both proteins were most active at pH 8.0. AbfA and Abf2 both belong to family 51 of the glycoside hydrolases but have different substrate specificity. AbfA acts preferentially on (1-->5) linkages of linear alpha-1,5-l-arabinan and alpha-1,5-linked arabino-oligomers, and is much less effective on branched sugar beet arabinan and arabinoxylan and arabinogalactan. In contrast, Abf2 is most active on (1-->2) and (1-->3) linkages of branched arabinan and arabinoxylan, suggesting a concerted contribution of these enzymes to optimal utilization of arabinose-containing polysaccharides by B. subtilis.This work was partially supported by grant no. POCI/AGR/60236/2004 from the Fundacao para a Ciencia e Tecnologia (FCT) and FEDER to I. d. S.- N., and fellowship SFRH/BD/18238/2004 from the FCT to J. M. I.SGMRUNSá-Nogueira, Isabel deInácio, José ManuelCorreia, Isabel Lopes2010-10-25T10:30:47Z20082008-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10362/4200engInácio, JM, Correia, I.L. and de Sá-Nogueira I. (2008). Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degradation in Bacillus subtilis. Microbiology-SGM 154 (9): 2719-2729.1350-0872 (Print)info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T03:33:53Zoai:run.unl.pt:10362/4200Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:15:35.595645Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degradation in Bacillus subtilis.
title Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degradation in Bacillus subtilis.
spellingShingle Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degradation in Bacillus subtilis.
Sá-Nogueira, Isabel de
title_short Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degradation in Bacillus subtilis.
title_full Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degradation in Bacillus subtilis.
title_fullStr Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degradation in Bacillus subtilis.
title_full_unstemmed Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degradation in Bacillus subtilis.
title_sort Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degradation in Bacillus subtilis.
author Sá-Nogueira, Isabel de
author_facet Sá-Nogueira, Isabel de
Inácio, José Manuel
Correia, Isabel Lopes
author_role author
author2 Inácio, José Manuel
Correia, Isabel Lopes
author2_role author
author
dc.contributor.none.fl_str_mv RUN
dc.contributor.author.fl_str_mv Sá-Nogueira, Isabel de
Inácio, José Manuel
Correia, Isabel Lopes
description Microbiology 154 (2008) 2719-2729
publishDate 2008
dc.date.none.fl_str_mv 2008
2008-01-01T00:00:00Z
2010-10-25T10:30:47Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10362/4200
url http://hdl.handle.net/10362/4200
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Inácio, JM, Correia, I.L. and de Sá-Nogueira I. (2008). Two distinct arabinofuranosidases contribute to arabino-oligosaccharide degradation in Bacillus subtilis. Microbiology-SGM 154 (9): 2719-2729.
1350-0872 (Print)
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